长江中下游稻区不同水旱轮作模式和氮肥水平 对稻田CH4排放的影响

刘少文,殷敏,褚光,徐春梅,王丹英,章秀福,陈松

Effects of Various Paddy-Upland Crop Rotations and Nitrogen Fertilizer Levels on CH4 Emission in the Middle and Lower Reaches of the Yangtze River

ShaoWen LIU,Min YIN,Guang CHU,ChunMei XU,DanYing WANG,XiuFu ZHANG,Song CHEN

表2 光实时定量PCR扩增引物及反应条件

Table 2 Aplification primer and reaction condition of quantitative PCR

目的基因 Target gene 引物 Primer 引物序列(5′→3′) Sequence (5′→3′) 定量PCR反应程序 Thermal profile mcrA[26] MLf GGTGGTGTMGGATTCACACARTAYGCWACAGC 94℃预变性3 min,94℃变性25 s,50℃退火45 s,72℃延伸60 s,35个循环 Pre-denaturation at 94℃ for 3 min, denaturation at 94℃ for 25 s, annealing at 50℃ for 45 s, extension at 72°C for 60 s, 35 cycles MLr TTCATTGCRTAGTTWGGRTAGTT pmoA[27] PmoA A189f GGNGACTGGGACTTCTGG 95℃预变性5 min,92℃变性1 min,55℃退火1.5 min,72℃延伸60 s,35个循环 Pre-denaturation at 95℃ for 5 min, denaturation at 92℃ for 1 min, annealing at 55℃ for 1.5 min, extension at 72℃ for 60 s, 35 cycles pmoA mb661r CCGGMGCAACGTCYTTACC